shRNA Knockdown of ILF3 in Monocyte-Derived Dendritic Cells (MDDCs)

Antigen presenting cells such as myeloid dendritic cells (DCs) are key sentinels of the innate immune system. In response to pathogen recognition and innate immune stimulation, DCs transition from an immature, inactive state to a mature, active state that is characterized by widespread changes in host gene expression. Several key transcription factors are known to drive these gene expression changes, but the mechanisms that negatively regulate DC maturation are less well understood. Here, we have identified that the transcription factor Interleukin Enhancer Binding Factor 3 (ILF3) is a negative regulator of innate immune responses and DC maturation. Depletion of ILF3 in primary human monocyte-derived DCs (MDDCs) using shRNA knockdown led to increased expression of maturation markers and potentiated innate responses at baseline. DCs were derived from the peripheral blood from three deidentified human donors and transduced with lentiviral control or two individual shRNAs targeting ILF3 in the presence of Vpx. RNA expression was measured at day 5 post-transduction.